CN109265758B - temperature/pH dual-response type chitin nanofiber hydrogel and preparation method thereof - Google Patents
temperature/pH dual-response type chitin nanofiber hydrogel and preparation method thereof Download PDFInfo
- Publication number
- CN109265758B CN109265758B CN201811067274.7A CN201811067274A CN109265758B CN 109265758 B CN109265758 B CN 109265758B CN 201811067274 A CN201811067274 A CN 201811067274A CN 109265758 B CN109265758 B CN 109265758B
- Authority
- CN
- China
- Prior art keywords
- temperature
- chitin
- dual
- hydrogel
- chitin nanofiber
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
- C08J3/075—Macromolecular gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
- C08J2305/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2403/00—Characterised by the use of starch, amylose or amylopectin or of their derivatives or degradation products
- C08J2403/02—Starch; Degradation products thereof, e.g. dextrin
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/14—Hemicellulose; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K5/00—Use of organic ingredients
- C08K5/16—Nitrogen-containing compounds
- C08K5/17—Amines; Quaternary ammonium compounds
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K5/00—Use of organic ingredients
- C08K5/49—Phosphorus-containing compounds
- C08K5/51—Phosphorus bound to oxygen
- C08K5/52—Phosphorus bound to oxygen only
- C08K5/521—Esters of phosphoric acids, e.g. of H3PO4
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dispersion Chemistry (AREA)
- Polymers & Plastics (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Inorganic Chemistry (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Preparation (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a temperature/pH dual-response hydrogel and a preparation method thereof, wherein the method comprises the following steps: preparing partial chitosan; preparing chitin nanofiber dispersion liquid; slowly dripping and mixing the water phase, the oil phase and the surfactant according to the proportion, and stirring continuously for 2-72 hours to obtain clear and transparent oil-in-water microemulsion; evenly mixing the chitin nano dispersion liquid, the microemulsion, the beta-glycerophosphate and the biological compound factor in proportion, and obtaining the temperature/pH dual-response hydrogel through an alkaline coagulation bath. The temperature/pH dual-response hydrogel is prepared by compounding the beta-GP, the biological compound factor and the chitin nano-fibers, and the preparation method is simple and has strong operability. The temperature/pH dual-response hydrogel has good mechanical property, biocompatibility, temperature responsiveness, pH responsiveness and slow release performance, can be applied to the medicine field of medicine slow release and the like, and has good application prospect.
Description
Technical Field
The invention relates to the field of nanofiber hydrogel preparation, in particular to temperature/pH dual-response chitin nanofiber hydrogel and a preparation method thereof.
Background
Chitin (Chitin) is a natural high molecular polymer connected by beta- (1-4) glycosidic bonds, and can be dispersed in weak acid solution after partial deacetylation treatment. The surface of the chitin fiber crystal has enough free amino groups and high-density positive charges under acidic conditions, and the nanofiber is obtained after certain mechanical treatment. The chitin nano-fiber has particularly excellent biological performance, and the sufficient number of amino groups on the surface of the chitin nano-fiber ensures that the chitin nano-fiber has better biological and chemical reaction performance, so that the chitin nano-fiber can be widely applied to biomedical products, drug delivery systems and the like. With the improvement of the requirement of people on the utilization rate of some expensive and insoluble drugs and the increasing attention on the slow release and controlled release characteristics of the drugs, the nano material hydrogel has wider application prospect as an embedded microemulsion type drug carrier. The close combination of the nanometer material field and the relevant fields such as biomedicine provides a brand new idea for medical treatment. The temperature/pH dual-response nanofiber hydrogel is the key point of research in the field of biomedicine, but the development of the temperature/pH dual-response nanofiber hydrogel is limited because the preparation process of the temperature/pH dual-response nanofiber hydrogel mainly relates to chemical grafting modification at present, the reaction process is complex, and uncontrollable factors are more.
Disclosure of Invention
The purpose of the invention is as follows: aiming at the defects in the prior art, the invention aims to provide a preparation method of temperature/pH dual-response type chitin nanofiber hydrogel, which can be prepared by simple physical compounding and biological factor compounding, and has simple preparation method and strong operability. Another object of the present invention is to provide a temperature/pH dual-response type chitin nanofiber hydrogel having excellent temperature/pH dual-response properties, biocompatibility and drug sustained-release properties,
the technical scheme is as follows: in order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of temperature/pH dual-response type chitin nanofiber hydrogel comprises the following steps:
(1) preparing partial chitosan;
(2) preparing chitin nanofiber dispersion liquid;
(3) slowly dripping and mixing the water phase, the oil phase and the surfactant according to a proportion, and stirring continuously for 2-72 hours to obtain clear and transparent oil-in-water (O/W) microemulsion;
(4) uniformly mixing the chitin nano dispersion liquid, the microemulsion, the beta-glycerophosphate (beta-GP) and the biological compound factor in proportion, and carrying out alkaline coagulation bath to obtain the temperature/pH dual-response chitin nanofiber hydrogel.
In the step (1), the chitin is at least one of the exoskeleton of crabs or shrimps, the parietal bone of squid, the cuticle of insects and the cell wall of fungi; the preparation method of chitosan comprises chemical deacetylation and biological enzymatic deacetylation.
In the step (2), the mass percentage concentration of the chitin nano-fiber dispersion liquid is 0.2-0.8%, and the medium is an acidic medium; wherein the acidic medium is at least one of acetic acid, gluconic acid, itaconic acid, citric acid, ascorbic acid, lactic acid, hydrochloric acid, phosphoric acid and sulfuric acid.
In the step (3), the particle size of the microemulsion is 2-100 nm, and the oil phase is at least one selected from isopropyl myristate, ethyl oleate, ethyl acetate, corn oil, olive oil, petroleum ether and lubricating oil; the surfactant is at least one of Tween series and Span series.
In the step (4), the beta-GP is prepared by beta-glycerophosphate or beta-glycerophosphate, and the preferred concentration range is 30-60% (w/w); the biological compound factor is at least one of chitosan, aspartic acid, serine, glycine, xylan and starch, preferably in the concentration range of 1-10% (w/w).
In the preparation method of the temperature/pH dual-response type chitin nanofiber hydrogel, the mass ratio of beta-GP to chitin nanofibers is 1: 0.4-5; the mass ratio of the biological compound factor to the chitin nano-fiber is 1: 0.2-3.
The temperature/pH dual-response type chitin nanofiber hydrogel prepared by the preparation method of the temperature/pH dual-response type hydrogel.
The temperature/pH dual-response type chitin nano-fiber hydrogel has a temperature response range of 20-50 ℃ and a pH response range of 1.5-7.4.
The temperature/pH dual-response type chitin nano-fiber hydrogel is applied to the preparation of a slow-release material.
The temperature/pH dual-response type chitin nano-fiber hydrogel is applied to the preparation of sustained-release drugs.
Has the advantages that: compared with the prior art, the invention has the advantages that: the chitin nanofiber hydrogel is compounded with beta-GP and biological compound factors to prepare the hydrogel with temperature sensitivity and pH sensitivity, so that the rehydration gel has excellent temperature sensitivity, the preparation method is simple, the preparation process is green and economic, and the operability is strong; the temperature/pH dual-response type chitin nanofiber hydrogel prepared by the invention has the advantages of environmental protection, good biocompatibility and drug slow release performance.
Drawings
FIG. 1 is a stress scan of chitin nanofiber dispersion after being compounded with beta-GP at different temperatures;
FIG. 2 is a graph of the mechanical properties of chitin nanofiber composite hydrogels;
FIG. 3 is a graph of citric acid dispersed chitin nanofiber composite hydrogel release;
FIG. 4 is a slow release curve chart of temperature-sensitive type gel of chitin nano-fiber added with glycine under the conditions of 25 ℃ and 40 ℃;
FIG. 5 is a slow release curve chart of chitosan-added chitin nanofiber temperature-sensitive gel at 25 ℃ and 40 ℃;
FIG. 6 is a slow release curve chart of temperature-sensitive type gel of chitin nano-fiber added with glycine starch at 25 ℃ and 40 DEG C
FIG. 7 is a graph of the sustained release of chitin nanofiber hydrogel at different pH and temperature conditions.
Detailed Description
The invention is further illustrated by the following examples. The examples are intended to illustrate, but not to limit, the invention. It will be understood by those of ordinary skill in the art that these examples are not intended to limit the present invention in any way and that appropriate modifications may be made without departing from the spirit and scope of the present invention.
Example 1
(1) Preparation of partially deacetylated chitin
Cutting crab shell into 1cm2And (3) soaking the materials for more than 12 hours in 1mol/L NaOH to remove protein, washing the materials to be neutral by distilled water, then soaking the materials for more than 12 hours in 1mol/L HCl to remove minerals, and washing the materials to be neutral. The above steps were repeated three times. Configuring 0.5% (w/w) of NaClO2Adjusting the pH value of the mixture to 5 with acetic acid, bleaching the mixture for 2 hours in water bath at 70 ℃, intermittently stirring the mixture, and washing the chitin to be neutral. Crushing with a juicer, filtering to remove water, storing in a refrigerator at 4 deg.C, and balancing water for 1-2 days.
1g of purified chitin (dry weight) was weighed, soaked in 50mL of 35% (w/w) NaOH solution, and subjected to partial deacetylation treatment by stirring in a water bath at 90 ℃ for 4 h. After the reaction was completed, the reaction mixture was washed to neutrality, and the solid was collected. Storing in a refrigerator at 4 deg.C, and balancing water.
(2) Preparation of chitin nanodispersion
Preparing a chitin/water mixed system with a mass ratio of 0.2% (w/w) in a 200mL beaker, dropwise adding an acetic acid solution into the chitin dispersion to maintain the pH of the solution at 3-4, fully swelling, and then homogenizing for 30s by a homogenizer and dispersing for 5min by ultrasound for five times respectively. Centrifuging at 10000rpm/min for 5min to remove undispersed solid, collecting supernatant, pouring into a reagent bottle, and collecting to obtain chitin nanometer dispersion.
(3) Preparation of additive liquid
Preparing a beta-glycerophosphate (beta-GP) solution: beta-GP was formulated from disodium beta-phosphate and was formulated as a 50% (w/w) solution and stored in a refrigerator at 4 ℃.
The biocomplex factor Chitosan (CS), aspartic acid (Asp), serine (Ser), glycine (Gly), Xylan (Xylan) and Starch (Starch) were prepared as 1.5% (w/w) solutions, respectively. Dissolving CS with acetic acid, heating and stirring; asp, Ser, Gly, Xyaln and Starch were dissolved in distilled water.
(4) Preparation of composite dual-response hydrogel
The chitin nano-fiber dispersion liquid with the concentration of 0.4 percent and the mixed biological compound factor solution with the concentration of 1.5 percent are mixed according to the proportion of 1: 1.5, the beta-GP and the microemulsion with the proportion of 1: 2 to the nano-fibers are added, and the mixture is uniformly mixed to prepare the hydrogel with temperature sensitivity and pH sensitivity under the condition of alkaline ammonia water bath.
(5) Sustained release of hydrogels
The hydrogel was slowly released from the buffer (pH 7.475mM) at a ratio of 2: 100(v/v), and 600uL of the sample was taken over a period of time and replaced with the same amount of fresh buffer, keeping the total volume of the sustained release constant. The conditions are as follows: the rotating speed is 30 +/-2 rpm/min, and the human body environment is simulated.
Example 2
(1) Preparation of partially deacetylated chitin
Soaking the squid parietal bone in 1mol/L NaOH for more than 12h to remove protein, washing with distilled water to neutrality, soaking with 1mol/L HCl for more than 12h to remove mineral substances, and washing to neutrality. The above steps were repeated three times. Configuring 0.5% (w/w) of NaClO2Adjusting the pH value of acetic acid to 5, bleaching in water bath at 70 ℃ for 2h, intermittently stirring, and washing the chitin to be neutral. Crushing with a juicer, filtering to remove water, storing in a refrigerator at 4 deg.C, and balancing water for 1-2 days.
1g of purified chitin (dry weight) was weighed, soaked in 50mL of 35% (w/w) NaOH solution, and subjected to partial deacetylation treatment by stirring in a water bath at 90 ℃ for 4 h. After the reaction was completed, the reaction mixture was washed to neutrality, and the solid was collected. Storing in a refrigerator at 4 deg.C, and balancing water.
(2) Chitin nanodispersion was prepared as in example 1
(3) Preparation of additive liquid
Preparing a beta-GP solution: the beta-GP is prepared from beta-calcium glycerophosphate, and the beta-GP is prepared into a 30% (w/w) solution and stored in a refrigerator at 4 ℃.
The biocomplex factor Chitosan (CS), aspartic acid (Asp), serine (Ser), glycine (Gly), Xylan (Xylan) and Starch (Starch) were prepared into 5% (w/w) solutions, respectively. Dissolving CS with acetic acid, heating and stirring; asp, Ser, Gly, Xyaln and Starch were dissolved in distilled water.
(4) Preparation of composite dual-response hydrogel
Mixing 0.6% chitin nanofiber dispersion liquid and 5% glycine solution according to a ratio of 1: 3, adding beta-GP and microemulsion according to a ratio of 1: 4 to the nanofibers, and uniformly mixing to prepare the hydrogel with temperature sensitivity and pH sensitivity under the condition of alkaline ammonia water bath.
(5) Sustained release of hydrogel was the same as in example 1
Example 3
(1) Preparation of partially deacetylated chitin
Soaking periostracum Cicadae in 1mol/L NaOH for more than 12 hr to remove protein, washing with distilled water to neutrality, soaking with 1mol/L HCl for more than 12 hr to remove mineral substance, and washing to neutrality. The above steps were repeated three times. Configuring 0.5% (w/w) of NaClO2Adjusting the pH value of the mixture to 5 with acetic acid, bleaching the mixture for 2 hours in water bath at 70 ℃, intermittently stirring the mixture, and washing the chitin to be neutral. Crushing with a juicer, filtering to remove water, storing in a refrigerator at 4 deg.C, and balancing water for 1-2 days.
1g of purified chitin (dry weight) was weighed, soaked in 50mL of 35% (w/w) NaOH solution, and subjected to partial deacetylation treatment by stirring in a water bath at 90 ℃ for 4 h. After the reaction was completed, the reaction mixture was washed to neutrality, and the solid was collected. Storing in a refrigerator at 4 deg.C, and balancing water.
(2) Preparation of chitin nanodispersion
Preparing a chitin/water mixed system with a mass ratio of 0.2% (w/w) in a 200mL beaker, dropwise adding a citric acid solution into the chitin dispersion liquid to maintain the pH of the solution at 3-4, fully swelling, and then homogenizing for 30s by a homogenizer and dispersing for 5min by ultrasound for five times respectively. Centrifuging at 10000rpm/min for 5min to remove undispersed solid, collecting supernatant, pouring into a reagent bottle, and collecting to obtain chitin nanometer dispersion.
(3) Preparation of additive liquid
Preparing a beta-glycerophosphate (beta-GP) solution: beta-GP was formulated from disodium beta-phosphate and was formulated as a 50% (w/w) solution and stored in a refrigerator at 4 ℃.
The biocomplex factor Chitosan (CS), aspartic acid (Asp), serine (Ser), glycine (Gly), Xylan (Xylan) and Starch (Starch) were prepared as 3.0% (w/w) solutions, respectively. Dissolving CS with acetic acid, heating and stirring; asp, Ser, Gly, Xyaln and Starch were dissolved in distilled water.
(4) Preparation of composite dual-response hydrogel
Mixing 0.4% chitin nano fiber dispersion solution and 3.0% xylan solution in the ratio of 1 to 1.5, adding beta-GP and microemulsion in the ratio of 1 to 1, and mixing uniformly to prepare the hydrogel with temperature sensitivity and pH sensitivity under the condition of alkaline ammonia water bath.
(5) Sustained release of hydrogel was the same as in example 1
Example 4
(1) Preparation of partially deacetylated chitin
Soaking shrimp shell in 1mol/L NaOH for more than 12h to remove protein, washing with distilled water to neutrality, soaking with 1mol/L HCl for more than 12h to remove mineral substances, and washing to neutrality. The above steps were repeated three times. Configuring 0.5% (w/w) of NaClO2Adjusting the pH value of the mixture to 5 with acetic acid, bleaching the mixture for 2 hours in water bath at 70 ℃, intermittently stirring the mixture, and washing the chitin to be neutral. Crushing with a juicer, filtering to remove water, storing in a refrigerator at 4 deg.C, and balancing water for 1-2 days.
Weighing 1g purified chitin (dry weight), soaking in buffer solution with pH of 9.0, adding chitin deacetylase with certain enzyme activity, and performing enzymatic deacetylation reaction at 50 deg.C for 48 h. The reaction is terminated, and the sample is washed to be neutral to obtain partial chitosan. Storing in refrigerator at 4 deg.C, measuring water content after 24 hr, and using as enzymatic partial chitosan wet sample.
(2) Preparation of chitin nanodispersion
Preparing a chitin/water mixed system with a mass ratio of 0.2% (w/w) in a 200mL beaker, dropwise adding a citric acid solution into the chitin dispersion liquid to maintain the pH of the solution at 3-4, fully swelling, and then homogenizing for 30s by a homogenizer and dispersing for 5min by ultrasound for five times respectively. Centrifuging at 10000rpm/min for 5min to remove undispersed solid, collecting supernatant, pouring into a reagent bottle, and collecting to obtain chitin nanometer dispersion.
(3) Preparation of additive liquid
Preparing a beta-glycerophosphate (beta-GP) solution: beta-GP was formulated from disodium beta-phosphate and was formulated as a 50% (w/w) solution and stored in a refrigerator at 4 ℃.
The biocomplex factor Chitosan (CS), aspartic acid (Asp), serine (Ser), glycine (Gly), Xylan (Xylan) and Starch (Starch) were prepared as 1.5% (w/w) solutions, respectively. Dissolving CS with acetic acid, heating and stirring; asp, Ser, Gly, Xyaln and Starch were dissolved in distilled water.
(4) Preparation of composite dual-response hydrogel
Mixing 0.4% chitin nano fiber dispersion solution and 1.5% xylan solution in the ratio of 1 to 1, adding beta-GP and microemulsion in the ratio of 1 to 1, and mixing uniformly to prepare the hydrogel with temperature sensitivity and pH sensitivity under the condition of alkaline ammonia water bath.
(5) Sustained release of hydrogel was the same as in example 1
Example 5
(1) Preparation of partially deacetylated chitin
Soaking shrimp shell in 1mol/L NaOH for more than 12h to remove protein, washing with distilled water to neutrality, soaking with 1mol/L HCl for more than 12h to remove mineral substances, and washing to neutrality. The above steps were repeated three times. Configuring 0.5% (w/w) of NaClO2Adjusting the pH value of the mixture to 5 with acetic acid, bleaching the mixture for 2 hours in water bath at 70 ℃, intermittently stirring the mixture, and washing the chitin to be neutral. Crushing with a juicer, filtering to remove water, storing in a refrigerator at 4 deg.C, and balancing water for 1-2 days.
Weighing 1g purified chitin (dry weight), soaking in buffer solution with pH of 9.0, adding chitin deacetylase with certain enzyme activity, and performing enzymatic deacetylation reaction at 50 deg.C for 48 h. The reaction is terminated, and the sample is washed to be neutral to obtain partial chitosan. Storing in refrigerator at 4 deg.C, measuring water content after 24 hr, and using as enzymatic partial chitosan wet sample.
(2) Preparation of chitin nanodispersion
Preparing a chitin/water mixed system with a mass ratio of 0.2% (w/w) in a 200mL beaker, dropwise adding a hydrochloric acid solution into the chitin dispersion liquid to maintain the pH of the solution at 3-4, fully swelling, and then homogenizing for 30s by a homogenizer and dispersing for 5min by ultrasound for five times respectively. Centrifuging at 10000rpm/min for 5min to remove undispersed solid, collecting supernatant, pouring into a reagent bottle, and collecting to obtain chitin nanometer dispersion.
(3) Preparation of additive liquid
Preparing a beta-glycerophosphate (beta-GP) solution: beta-GP was formulated from disodium beta-phosphate and was formulated as a 50% (w/w) solution and stored in a refrigerator at 4 ℃.
The biocomplex factor Chitosan (CS), aspartic acid (Asp), serine (Ser), glycine (Gly), Xylan (Xylan) and Starch (Starch) were prepared as 1.5% (w/w) solutions, respectively. Dissolving CS with acetic acid, heating and stirring; asp, Ser, Gly, Xyaln and Starch were dissolved in distilled water.
(4) Preparation of composite dual-response hydrogel
Mixing 0.3% chitin nano fiber dispersion solution and 1.5% xylan and serine mixed solution according to the proportion of 1: 1, adding beta-GP and microemulsion according to the proportion of 1: 1 to the nano fibers, and uniformly mixing to prepare the hydrogel with temperature sensitivity and pH sensitivity under the condition of alkaline ammonia water bath.
(5) Sustained release of hydrogel was the same as in example 1
Example 6
(1) Partial chitosan was prepared as in example 1;
(2) the chitin nanodispersion was prepared as in example 1;
(3) preparation of additive liquid
Preparing a beta-glycerophosphate (beta-GP) solution: beta-GP was formulated from calcium glycerophosphate beta-GP as a 50% (w/w) solution and stored in a refrigerator at 4 ℃.
The biocomplex factor Chitosan (CS), aspartic acid (Asp), serine (Ser), glycine (Gly), Xylan (Xylan) and Starch (Starch) were prepared as 1.5% (w/w) solutions, respectively. Dissolving CS with acetic acid, heating and stirring; asp, Ser, Gly, Xyaln and Starch were dissolved in distilled water.
(4) Preparation of composite dual-response hydrogel
The chitin nano-fiber dispersion liquid with the concentration of 0.5 percent and the aspartic acid solution with the concentration of 1.5 percent are mixed according to the proportion of 1: 1, the beta-GP and the microemulsion with the proportion of 1: 1 to the nano-fibers are added and uniformly mixed to prepare the hydrogel with temperature sensitivity and pH sensitivity under the condition of alkaline ammonia water bath.
(5) Sustained release of hydrogel was the same as in example 1
Example 7
(1) Preparation of partially deacetylated chitin
Soaking crab shell in 1mol/L NaOH for more than 12h to remove protein, washing with distilled water to neutrality, soaking with 1mol/L HCl for more than 12h to remove mineral substance, and washing to neutrality. The above steps were repeated three times. Configuring 0.5% (w/w) of NaClO2Adjusting the pH value of the mixture to 5 with acetic acid, bleaching the mixture for 2 hours in water bath at 70 ℃, intermittently stirring the mixture, and washing the chitin to be neutral. Crushing with a juicer, filtering to remove water, storing in a refrigerator at 4 deg.C, and balancing water for 1-2 days.
Weighing 1g purified chitin (dry weight), soaking in buffer solution with pH of 9.0, adding chitin deacetylase with certain enzyme activity, and performing enzymatic deacetylation reaction at 50 deg.C for 48 h. The reaction is terminated, and the sample is washed to be neutral to obtain partial chitosan. Storing in refrigerator at 4 deg.C, measuring water content after 24 hr, and using as enzymatic partial chitosan wet sample.
(2) Chitin nanodispersion was prepared as in example 1
(3) Preparation of additive liquid
Preparing a beta-glycerophosphate (beta-GP) solution: the beta-GP is prepared from beta-calcium glycerophosphate, and the beta-GP is prepared into a 60% (w/w) solution and stored in a refrigerator at 4 ℃.
The biocomplex factor Chitosan (CS), aspartic acid (Asp), serine (Ser), glycine (Gly), Xylan (Xylan) and Starch (Starch) were prepared as 2.0% (w/w) solutions, respectively. Dissolving CS with acetic acid, heating and stirring; asp, Ser, Gly, Xyaln and Starch were dissolved in distilled water.
(4) Preparation of composite dual-response hydrogel
Mixing 0.5% chitin nano fiber dispersion solution and 2.0% starch solution according to a ratio of 1: 1, adding beta-GP and microemulsion according to a ratio of 1: 1 to the nano fibers, and uniformly mixing to prepare the hydrogel with temperature sensitivity and pH sensitivity under the condition of alkaline ammonia water bath.
(5) Sustained release of hydrogel was the same as in example 1
Example 8
The performance of the hydrogels prepared in examples 1 to 7 was examined as follows:
fig. 1 is a stress scan of the chitin nanofiber dispersion liquid compounded with β -GP at different temperatures, and it can be seen that the β -GP-compounded chitin nanofiber dispersion liquid shows liquid properties at the previous stage, and changes into solid properties after passing through an intersection point, and the phase angle corresponding to the intersection point is 35 ℃.
FIG. 2 is a graph of the mechanical properties of chitin nanofiber composite hydrogel, and it can be seen that the addition of β -GP affects the mechanical properties of the composite gel.
Fig. 3 is a sustained-release graph of a chitosan nanofiber composite hydrogel dispersed with citric acid, and it can be seen that the sustained-release amount of the hydrogel prepared under the same conditions is different by 40% under two temperature conditions, which also indicates that the addition of β -GP helps to improve the temperature sensitivity of the nanofiber hydrogel.
Fig. 4-6 are sustained-release graphs of chitin nanofiber hydrogels compounded by different biocomposite factors, and the compounded chitin nanofiber hydrogels showed good temperature sensitivity.
Fig. 7 is a sustained-release diagram of the chitin nanofiber hydrogel under different pH and temperature conditions, which shows that the composite hydrogel has excellent temperature/pH dual responsiveness.
Claims (8)
1. A preparation method of temperature/pH dual-response type chitin nanofiber hydrogel is characterized by comprising the following steps:
(1) preparing partial chitosan;
(2) preparing chitin nanofiber dispersion liquid;
(3) slowly dripping and mixing the water phase, the oil phase and the surfactant according to a proportion, and stirring continuously for 2-72 hours to obtain clear and transparent oil-in-water microemulsion;
(4) uniformly mixing the chitin nanofiber dispersion liquid, the microemulsion, the beta-glycerophosphate and the biological compound factor in proportion, and carrying out alkaline coagulation bath to obtain a temperature/pH dual-response type chitin nanofiber hydrogel; wherein the biological compound factor is at least one of chitosan, aspartic acid, serine, glycine, xylan and starch; wherein, the mass percentage concentration of the chitin nano-fiber dispersion liquid is 0.2-0.8%, the mass percentage concentration of the beta-glycerophosphate solution is 30-60%, and the mass percentage concentration of the biological compound factor solution is 1-10%; the mass ratio of the beta-glycerophosphate solution to the chitin nanofiber solution is 1: 0.4-5; the mass ratio of the biological compound factor solution to the chitin nano-fiber solution is 1: 0.2-3.
2. The method for preparing a temperature/pH bi-responsive chitin nanofiber hydrogel according to claim 1, wherein in the step (1), the chitin is derived from at least one of exoskeletons of crabs or shrimps, parietal bones of squids, cuticles of insects, and cell walls of fungi; the chitosan is prepared by chemical deacetylation and/or biological enzymatic deacetylation.
3. The method for preparing temperature/pH dual-response type chitin nanofiber hydrogel according to claim 1, wherein in the step (2), the mass percentage concentration of the chitin nanofiber dispersion is 0.2-0.8%, and the medium is an acidic medium; wherein the acidic medium is at least one of acetic acid, gluconic acid, itaconic acid, citric acid, ascorbic acid, lactic acid, hydrochloric acid, phosphoric acid and sulfuric acid.
4. The method for preparing temperature/pH dual-response type chitin nanofiber hydrogel according to claim 1, wherein in the step (3), the particle size of the microemulsion is 2-100 nm, and the oil phase is at least one selected from isopropyl myristate, ethyl oleate, ethyl acetate, corn oil, olive oil, petroleum ether and lubricating oil; the surfactant is at least one of Tween series and Span series.
5. The temperature/pH dual-response type chitin nanofiber hydrogel prepared by the method for preparing the temperature/pH dual-response type hydrogel according to any one of claims 1 to 4.
6. The temperature/pH dual-response chitin nanofiber hydrogel according to claim 5, wherein the temperature response range of said hydrogel is 20-50 ℃ and the pH response range is 1.5-7.4.
7. The use of the temperature/pH bi-responsive chitin nanofiber hydrogel of claim 5 in the preparation of a sustained release material.
8. The use of the temperature/pH bi-responsive chitin nanofiber hydrogel of claim 5 in the preparation of a sustained release drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811067274.7A CN109265758B (en) | 2018-09-12 | 2018-09-12 | temperature/pH dual-response type chitin nanofiber hydrogel and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811067274.7A CN109265758B (en) | 2018-09-12 | 2018-09-12 | temperature/pH dual-response type chitin nanofiber hydrogel and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109265758A CN109265758A (en) | 2019-01-25 |
| CN109265758B true CN109265758B (en) | 2020-12-04 |
Family
ID=65189327
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811067274.7A Active CN109265758B (en) | 2018-09-12 | 2018-09-12 | temperature/pH dual-response type chitin nanofiber hydrogel and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109265758B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112940287B (en) * | 2021-02-03 | 2022-05-13 | 武汉大学 | Shape memory chitosan hydrogel and preparation method thereof |
| CN113244453B (en) * | 2021-04-16 | 2022-04-19 | 北京科技大学 | Preparation method and application of controllable multi-stage crosslinking injectable thermotropic phase-change hydrogel |
| CN113750076B (en) * | 2021-09-30 | 2022-11-25 | 大连理工大学 | Chitin-gelatin-based composite porous microspheres as drug carrier and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103357062A (en) * | 2012-03-26 | 2013-10-23 | 约泰实业股份有限公司 | Fibrous hydrogel and preparation method thereof |
| WO2017135498A1 (en) * | 2016-02-05 | 2017-08-10 | 주식회사 파마리서치프로덕트 | Temperature sensitive hydrogel composition including nucleic acid and chitosan |
| CN107050523A (en) * | 2016-12-23 | 2017-08-18 | 江南大学 | A kind of preparation method of new β tricalcium phosphates/chitosan composite bionic hydrogel |
| CN108192113A (en) * | 2018-01-09 | 2018-06-22 | 南京林业大学 | A kind of method of the preparation and its redisperse of highly concentrated hygrometric state and dry state chitin nanofiber/whisker |
-
2018
- 2018-09-12 CN CN201811067274.7A patent/CN109265758B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103357062A (en) * | 2012-03-26 | 2013-10-23 | 约泰实业股份有限公司 | Fibrous hydrogel and preparation method thereof |
| WO2017135498A1 (en) * | 2016-02-05 | 2017-08-10 | 주식회사 파마리서치프로덕트 | Temperature sensitive hydrogel composition including nucleic acid and chitosan |
| CN107050523A (en) * | 2016-12-23 | 2017-08-18 | 江南大学 | A kind of preparation method of new β tricalcium phosphates/chitosan composite bionic hydrogel |
| CN108192113A (en) * | 2018-01-09 | 2018-06-22 | 南京林业大学 | A kind of method of the preparation and its redisperse of highly concentrated hygrometric state and dry state chitin nanofiber/whisker |
Non-Patent Citations (4)
| Title |
|---|
| Rheological characterization of new thermosensitive hydrogels formed by chitosan, glycerophosphate, and phosphorylated β-cyclodextrin;Carlos Alberto Ramírez Barragán等;《Carbohydrate Polymers》;20180820;第471-481页 * |
| Stimuli-responsive chitosan-starch injectable hydrogels combined with encapsulated adipose-derived stromal cells for articular cartilage regeneration;Helena Sa-Lima等;《Soft Matter》;20101231;第5184-5195页 * |
| Temperature and pH-sensitive chitosan hydrogels: DSC,rheological and swelling evidence of a volume phase transition;F.M.Goycoole等;《Polymer Bulletin》;20060627;第225-234页 * |
| 几丁质纳米纤维/壳聚糖复合气凝胶微球的制备与表征;刘亮 等;《南京工业大学学报（自然科学版）》;20160331;第38卷(第2期);第51-55页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109265758A (en) | 2019-01-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109265758B (en) | temperature/pH dual-response type chitin nanofiber hydrogel and preparation method thereof | |
| Muzzarelli et al. | Emerging biomedical applications of nano-chitins and nano-chitosans obtained via advanced eco-friendly technologies from marine resources | |
| Elieh-Ali-Komi et al. | Chitin and chitosan: production and application of versatile biomedical nanomaterials | |
| Yeul et al. | Unprecedented chitin and chitosan: A chemical overview | |
| Mathur et al. | Chitin and chitosan, versatile polysaccharides from marine animals | |
| Younes et al. | Chitin and chitosan preparation from marine sources. Structure, properties and applications | |
| CN105820352B (en) | A kind of preparation method of electropositive chitin nanofiber hydrogels and aeroge | |
| Ang-atikarnkul et al. | Fabrication of cellulose nanofiber/chitin whisker/silk sericin bionanocomposite sponges and characterizations of their physical and biological properties | |
| CN106220867A (en) | A kind of elecrtonegativity chitin nanofiber hydrogels and the preparation method of aeroge | |
| JP2005529191A (en) | Cell wall derivatives from biomass and their preparation | |
| Horue et al. | Bacterial cellulose-based materials as dressings for wound healing | |
| Silva et al. | Bacterial cellulose and ECM hydrogels: An innovative approach for cardiovascular regenerative medicine | |
| Saravanan et al. | A review on extraction of polysaccharides from crustacean wastes and their environmental applications | |
| CN103319739A (en) | Preparation method of chitin nanofiber/polylactic acid composite film | |
| EP0977780B1 (en) | Hetero-polysaccharide conjugates, s-inp polysaccharide gels and methods of making and using the same | |
| Zhao et al. | Degradable porous carboxymethyl chitin hemostatic microspheres | |
| CN102344686A (en) | Method for preparing fibroin nanoparticles taking polyvinyl alcohol as stabilizer | |
| CN104784744A (en) | Compound method for preparing wound dressing by utilizing human hair keratin | |
| Arfin et al. | Bio-based materials: past to future | |
| CN107028872A (en) | A kind of composite aquogel, its preparation method and application | |
| Das et al. | A review on bio-polymers derived from animal sources with special reference to their potential applications | |
| Sun et al. | The recent progress of the cellulose-based antibacterial hydrogel | |
| CN103768654B (en) | Water-soluble fibroin powder and preparation method thereof | |
| CN105820267B (en) | A kind of skin wound preparation for repairing and its preparation method and application | |
| CN102573937B (en) | Degradable composite material containing chitin or chitosan |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |